INTRODUCTION:Pharmacogenetic (PGx) screening is intended to optimize drug efficacy and reduce adverse drug reactions. Current screening options include genotyping assays for preselected PGx variants and broader next-generation sequencing panels (NGS). Few studies have directly compared preemptive PGx screening methods. MATERIALS AND METHODS:The two PGx methods were compared in a cross-sectional study of adult Military Health System (MHS) clinic beneficiaries. Participants had initial targeted CYP2C19/CYP2D6 genotyping at a Military Health System Laboratory. Genotyping was followed by multi-gene NGS testing. Current prescriptions were recorded and potential drug-drug interactions screened to evaluate prescribing risk. RESULTS:All participants (100%) had at least one clinically actionable NGS panel result compared to 81% with targeted CYP2C19/CYP2D6 genotyping. Participants (n = 162) had an average of 6.6 (range 0-22) prescriptions and 2.7 (range 0-24) drug-drug interactions. Among those with at least one clinically actionable NGS result, 42% were currently taking medication with actionable CPIC guidelines (Level A/B), compared with 24% with CYP2C19/CYP2D6 genotyping. Sixteen participants (10%) had uncertain NGS panel results, with none for CYP2C19/CYP2D6 genotyping. CONCLUSIONS:Preemptive multi-gene NGS detected more clinically actionable PGx results than targeted CYP2C19/CYP2D6 genotyping. Effective PGx screening in the MHS may decrease preventable adverse effects and improve military readiness.

PURPOSE:The Mayo-Baylor RIGHT 10K Study enabled preemptive, sequence-based pharmacogenomics (PGx)-driven drug prescribing practices in routine clinical care within a large cohort. We also generated the tools and resources necessary for clinical PGx implementation and identified challenges that need to be overcome. Furthermore, we measured the frequency of both common genetic variation for which clinical guidelines already exist and rare variation that could be detected by DNA sequencing, rather than genotyping.METHODS:Targeted oligonucleotide-capture sequencing of 77 pharmacogenes was performed using DNA from 10,077 consented Mayo Clinic Biobank volunteers. The resulting predicted drug response-related phenotypes for 13 genes, including CYP2D6 and HLA, affecting 21 drug-gene pairs, were deposited preemptively in the Mayo electronic health record.RESULTS:For the 13 pharmacogenes of interest, the genomes of 79% of participants carried clinically actionable variants in 3 or more genes, and DNA sequencing identified an average of 3.3 additional conservatively predicted deleterious variants that would not have been evident using genotyping.CONCLUSION:Implementation of preemptive rather than reactive and sequence-based rather than genotype-based PGx prescribing revealed nearly universal patient applicability and required integrated institution-wide resources to fully realize individualized drug therapy and to show more efficient use of health care resources.

Aim: Pharmacogenomics (PGx) tests are performed on whole-blood or saliva specimens. In patients with a transplanted liver, PGx results may be discordant with hepatic drug metabolizing enzyme activity. We evaluate the incidence and impact of PGx testing in liver transplant recipients, detail potential errors and describe clinical decision support (CDS) solution implemented. Materials & methods: A retrospective cohort study of liver transplant recipients at Mayo Clinic who underwent PGx testing between 1 January 1996 and 7 October 2019 were characterized. Impact of a CDS solution was evaluated. Results: There were 129 PGx tests in 117 patients. PGx testing incidence increased before (per year incidence rate ratio = 1.45, 95% CI: 1.20-1.74, p < 0.001) and after transplant (incidence rate ratio = 1.48, 95% CI: 1.27-1.72, p < 0.001). Three erroneous PGx tests were avoided 6 months following CDS implementation. Conclusion: Incidence of PGx testing in liver transplant recipients is increasing, leading to erroneous therapeutic decisions. CDS interventions and education are needed to prevent errors.

Background Several small studies have previously investigated associations between the cytochrome P450 2D6 (CYP2D6) metabolism and response to opioids. We used a large sample of patients to study associations between CYP2D6 phenotypes and estimated CYP2D6 enzymatic activity scores with pain control and adverse reactions related to codeine and tramadol use. We conducted additional analyses to determine whether our results were consistent among men and women. Methods We used data from 2,877 participants in the RIGHT Protocol who were prescribed codeine and/or tramadol between 01/01/2005 and 12/31/2017 and who were not prescribed CYP2D6 inhibitors within 1 year prior to the opioid prescription. CYP2D6 phenotype categories were condensed into four groups: (1) Ultra-rapid and Rapid (n = 61), (2) Normal and Intermediate to Normal (n = 1,448), (3) Intermediate and Intermediate to Poor (n = 1,175), and (4) Poor metabolizer status (n = 193). Opioid-related outcomes included indications of poor pain control or adverse reactions related to medication use. We modeled the risk of each outcome using logistic regression, adjusting for age, sex, race, and ethnicity. Results The results revealed a trend from poor to ultra-rapid and rapid CYP2D6 phenotypes in which the risk of adverse reactions incrementally increased and the risk of poor pain control incrementally decreased. This trend reached statistical significance among female (but not male) participants. Among normal and intermediate to normal metabolizers, a larger proportion of women experienced adverse reactions relative to men. Discussion We replicated and extended the findings of previous research indicating associations between CYP2D6 phenotypes and response to opioids. In addition, the observed associations were stronger in women than in men. We recommend sex differences to be factored in future research investigating associations between pharmacogenomics and response to medications.

CYP2C9 and CYP2C19 are highly polymorphic pharmacogenes; however, clinically actionable genetic variability in drug metabolism due to these genes has been limited to a few common alleles. The identification and functional characterization of less-common open reading frame sequence variation might help to individualize therapy with drugs that are substrates for the enzymes encoded by these genes. The present study identified seven uncharacterized variants each in CYP2C9 and CYP2C19 using next-generation sequence data for 1013 subjects, and functionally characterized the encoded proteins. Constructs were created and transiently expressed in COS-1 cells for the assay of protein concentration and enzyme activities using fluorometric substrates and liquid chromatography-tandem mass spectrometry with tolbutamide (CYP2C9) and (S)-mephenytoin (CYP2C19) as prototypic substrates. The results were compared with the SIFT, Polyphen, and Provean functional prediction software programs. Cytochrome P450 oxidoreductase (CPR) activities were also determined. Positive correlations were observed between protein content and fluorometric enzyme activity for variants of CYP2C9 (P < 0.05) and CYP2C19 (P < 0.0005). However, CYP2C9 709G>C and CYP2C19 65A>G activities were much lower than predicted based on protein content. Substrate intrinsic clearance values for CYP2C9 218C>T, 343A>C, and CYP2C19 337G>A, 518C>T, 556C>T, and 557G>A were less than 25% of wild-type allozymes. CPR activity levels were similar for all variants. In summary, sequencing of CYP2C9 and CYP2C19 in 1013 subjects identified low-frequency variants that had not previously been functionally characterized. In silico predictions were not always consistent with functional assay results. These observations emphasize the need for high-throughput methods for pharmacogene variant mutagenesis and functional characterization.

BACKGROUND:Cytochrome P450 2D6 (CYP2D6) is responsible for the metabolism of clinically used drugs and other environmental exposures, but it is unclear whether the CYP2D6 phenotype is associated with adverse health outcomes. The aim was to determine the association of CYP2D6 phenotype with the risk of hospitalization or an emergency department (ED) visit among a group of primary care patients.METHODS:In this study, 929 adult patients underwent CYP2D6 testing. The primary outcome was risk of hospitalization or an ED visit from January 2005 through September 2014. CYP2D6 genotypes were interpreted as 1 of 7 clinical phenotypes, from ultrarapid to poor metabolizer, and patients with the extensive metabolizer phenotype were used as the reference group. The hazard ratios (HRs) and 95% confidence intervals (CIs) were estimated for finding the association of CYP2D6 phenotypes with the risk of hospitalization or an ED visit by using Cox proportional hazard models and adjusting for age and sex.RESULTS:The median age was 49 years (interquartile range, 46-52 years); 74% of patients had 3 or fewer chronic conditions, 285 had at least 1 hospitalization, and 496 had at least 1 ED visit. The risk of hospitalization was higher among patients who were ultrarapid metabolizers compared to extensive metabolizers (47% vs 30%; HR, 1.69; 95% CI, 1.11-2.57), as was the risk of an ED visit (62% vs 49%; HR, 1.50; 95% CI, 1.05-2.14). For poor metabolizers compared to extensive metabolizers, there was no difference in the risk of hospitalization (HR, 0.95; 95% CI, 0.58-1.56), but there was an increase in the risk of an ED visit (HR, 1.38; 95% CI, 0.96-1.98) (the difference was not statistically significant).CONCLUSION:We found an increased risk of hospitalization or an ED visit among ultrarapid compared to extensive CYP2D6 metabolizers. Further research identifying the mechanisms of the association and ultimate clinical utility is warranted.

BACKGROUND:Through allele specific PCR we studied 220 CYP2C19 compound heterozygous samples, of unknown ethnicity, to determine the haplotype for each of the variations within a sample.MATERIALS & METHODS:The genotypes assessed were: 180 *2 and *17 samples (100% in trans); 20 *2 and *11 samples (100% in cis); ten *4 and *17 samples (50% of the samples were *1/*4B and 50% *4A/*17); six *2, *11 and *17 samples (100% showed *2 and *11 in cis, and *17 in trans); two *2, *4 and *17 samples (100% *4B with *2 in trans); one sample with *17 and *34 (these were in trans); and one sample that contained *2, *17, c.463G>T (p.E155X; *17 and c.463G>T were in cis, with *2 in trans).RESULTS & CONCLUSION:In our study, we observed a different frequency for the *4B allele (when a sample contains both *4 and *17); and identified *17 occurring in cis with a novel nonsense allele. Accurately assessing a patient's genotype, including assignment of a haplotype, can be important when making a phenotype prediction.

Background: The triallelic serotonin transporter gene linked polymorphic region (5-HTTLPR) has been associated with alterations in thermal pain perception. The primary aim of this study was to investigate the associations between heat pain (HP) perception and the triallelic 5-HTTLPR in a large cohort of adults with chronic pain.Methods: The cohort included 277 adults with chronic pain who met inclusion criteria, and were consecutively admitted to an outpatient pain rehabilitation program from March 2009 through March 2010. Individuals were genotyped for the triallelic 5-HTTLPR (including rs25531) and categorized as high, intermediate, or low expressors of the serotonin transporter. Standardized measures of HP perception were obtained using a validated quantitative sensory test method of levels.Results: The distribution of the high, intermediate, and low expressing genotypes was 61 (22%), 149 (54%) and 67 (24%), respectively. The Hardy-Weinberg P-value was 0.204 which indicated no departure from equilibrium. A significant effect of genotype was observed for values of HP threshold (P = 0.029). Individual group comparisons showed that values of HP threshold were significantly greater in the intermediate compared to the high expressing group (P = 0.009) but not the low expressing group (P > 0.1). In a multiple variable linear regression model, the intermediate group (P = 0.034) and male sex (P = 0.021) were associated with significantly greater values of HP 0.5, but no significant genotype-by-sex interaction effect was observed.Conclusions: In this study that involved adults with chronic pain, the intermediate triallelic 5-HTTLPR expressing group, but not the low expressing group, was associated with greater HP thresholds compared to the high expressing group.

Cytochrome P450 2D6 (CYP2D6) is highly polymorphic. CYP2D6-2D7 hybrid genes can be present in samples containing CYP2D6*4 and CYP2D6*10 alleles. CYP2D7-2D6 hybrid genes can be present in samples with duplication signals and in samples with homozygous genotyping results. The frequency of hybrid genes in clinical samples is unknown. We evaluated 1390 samples for undetected hybrid genes by polymerase chain reaction (PCR) amplification, PCR fragment analysis, TaqMan copy number assays, DNA sequencing, and allele-specific primer extension assay. Of 508 CYP2D6*4-containing samples, 109 (21.5%) harbored CYP2D6*68 + *4-like, whereas 9 (1.8%) harbored CYP2D6*4N + *4-like. Of 209 CYP2D6*10-containing samples, 44 (21.1%) were found to have CYP2D6*36 + *10. Of 332 homozygous samples, 4 (1.2%) harbored a single CYP2D7-2D6 hybrid, and of 341 samples with duplication signals, 25 (7.3%) harbored an undetected CYP2D7-2D6 hybrid. Phenotype before and after accurate genotyping was predicted using a method in clinical use. The presence of hybrid genes had no effect on the phenotype prediction of CYP2D6*4- and CYP2D6*10-containing samples. Four of four (100%) homozygous samples containing a CYP2D7-2D6 gene had a change in predicted phenotype, and 23 of 25 (92%) samples with a duplication signal and a CYP2D7-2D6 gene had a change in predicted phenotype. Four novel genes were identified (CYP2D6*13A1 variants 1 and 2, CYP2D6*13G1, and CYP2D6*13G2), and two novel hybrid tandem structures consisting of CYP2D6*13B + *68×2 + *4-like and CYP2D6*13A1 variant 2 + *1×N were observed.

CYP2D6 is genotyped clinically for prediction of response to tamoxifen, psychotropic drugs and other medications. Phenotype prediction is dependent upon accurate genotyping. The CYP Allele Nomenclature Committee maintains the allelic nomenclature for CYP2D6; however, in some cases, the list of polymorphisms associated with a given allele is incomplete. Clinical laboratories and in vitro diagnostic manufacturers rely upon this nomenclature, in addition to the literature, to infer allelic function and haplotypes and when they design CYP2D6-testing platforms. This article provides more complete sequencing data for the CYP2D6*11 allele and describes the difficulties encountered in genotyping CYP2D6 when incomplete data are available. The CYP Allele Nomenclature Committee should provide clear information about the completeness of the original data used to define each allele.

Narcolepsy with cataplexy is a debilitating sleep disorder with an estimated prevalence of about 0.05%. Narcolepsy is caused by a selective loss of hypocretin (orexin) producing neurons in the perifornical hypothalamus. Based on the very strong association with the HLA subtype DQB1*0602, it is currently hypothesized narcolepsy is caused by an autoimmune-mediated process directed at the hypocretin neurons. So far however, studies focusing on general markers of (auto)immune activation, as well as humoral immunity against the hypocretin system have not yielded consistent results supporting this hypothesis.

Most studies of trends in antidepressant pharmacotherapy have focused on relatively mildly ill, nonpyschotic outpatients. In this report, we provide detailed information on psychotropic use among patients with unipolar depression participating in a large, multisite electroconculsive therapy (ECT) study. Adequacy of antidepressant medication trials was assessed with the Antidepressant Treatment History Form. Among patients with nonpsychotic depression, 27% (60/220) had not had an adequate trial of an antidepressant before ECT, and 63% (139/220) had had at least one inadequate trial. Surprisingly, 33% (79/243) of nonpsychotic patients had been prescribed an antipsychotic. Among patients with psychotic depression, 95% (101/106) had not been given an adequate combination of an antidepressant and antipsychotic agent, mostly due to low doses of the latter class. Among all patients in the trial, 61% (213/352) had been prescribed at least one benzodiazepine, and only 7% (24/352) had been given a lithium augmentation trial. Use of hypnotic agents and anticonvulsants was common. In conclusion, patients with severe depression referred for ECT with a unipolar depressive episode have high rates of psychotropic usage, much of which is inadequate.

Vascular diseases and asthma are two common conditions associated with considerable morbidity and mortality. Both are chronic, each has an inflammatory component, both are accompanied by remodelling of hollow tubes and both are amenable to early intervention. Those working in cardiovascular diseases have already made significant advances in terms of understanding the pathophysiology and the need to conduct large scale multicentre trials to address simple but pivotal questions about altering outcomes. Those of us who work in the respiratory field lag behind. A meeting was held in the Napa Valley, California, in May 1993, to bring together basic scientists and clinicians from each of these research fields, in order to address the similarities and differences between asthma and vascular diseases. The meeting was challenging, and thought provoking, and highlighted a number of similarities between asthma and vascular diseases. It also left those of us working in the respiratory field with a major worry - how little we know about the lesion in asthma. The meeting was held over two days. The first day was devoted to basic science, and the second to therapeutic intervention and the design of large multicentre trials to consider the impact of treatment on disease progression. We considered (in both diseases) aspects of the pathological changes and the mechanisms contributing to the remodelling which occurs. These were addressed in studies carried out using both in vivo and in vitro experimental techniques. In particular, the regulation of growth of airway and vascular smooth muscle, in addition to the role this plays in altered in vivo and in vitro reactivity was discussed. We summarized the similarities and differences between asthma and vascular disease (table 1). On the second day, we considered the effect of specific therapeutic intervention in asthma and the effects of this on structural changes. A particular highlight was the final presentation by Gresele on the considerable progress made by those in the cardiovascular area in the design and successful execution of large multicentre clinical trials, which posed very simple questions to huge numbers of patients. Our cardiovascular colleagues expressed some dismay over the fact that we in the respiratory field could not seem to agree on a definition of asthma, or which outcome variables were most appropriate to measure. In table 2, the parameters used in the International Study of Infarct Survival (ISIS) trials are shown, together with some suggestions for the design of the ideal comparable trial in asthma patients. One particularly useful example of the need for researchers in the respiratory field to follow developments in the cardiovascular area has arisen since the meeting. In a recent publication by O'BRIEN et al. [2] the current approaches to therapeutic intervention in atherosclerotic coronary arteries have been questioned. This study shows that

Recent epidemiologic studies have indicated that obsessive compulsive disorder is a fairly common psychiatric condition. In this article, obsessive compulsive disorder, its subtypes, and epidemiologic features are described. The common obsessions and compulsions are discussed, as are the comorbid conditions. Currently, etiologic hypotheses revolve around serotonergic and dopaminergic neurotransmitter systems. Management of patients with obsessive compulsive disorder involves education, pharmacotherapy, and behavior therapy, and recent advances in such treatments offer patients hope. Psychotherapy has not proved useful. In refractory cases, psychosurgical intervention remains an option.

Recent epidemiologic studies have indicated that obsessive compulsive disorder is a fairly common psychiatric condition. In this article, obsessive compulsive disorder, its subtypes, and epidemiologic features are described. The common obsessions and compulsions are discussed, as are the comorbid conditions. Currently, etiologic hypotheses revolve around serotonergic and dopaminergic neuro­ transmitter systems. Management of patients with obsessive compulsive disorder involves education, pharmacotherapy, and behavior therapy, and recent advances in such treatments offer patients hope. Psychotherapy has not proved useful. In refractory cases, psychosurgical intervention remains an option.

Antipsychotic agents have many indications and are frequently used. All physicians, regardless of their subspecialty, will most likely treat patients who receive these drugs. This article is designed to help nonpsychiatric physicians use antipsychotics appropriately. The indications, recommended dosages, side effects, and drug interactions of antipsychotic medications are reviewed. Receptor binding data are used to help predict the side-effect profile of these agents. Knowledge of these side effects and of the patient's medical condition helps the physician select a drug that the patient can tolerate. Management of overdose situations is discussed.

Between 1960 and 1979 528 patients with abdominal aortic aneurysms presented to the university department of surgery. Of these, 222 (42%) were elective cases, 72 acute (14%), 174 had ruptured (33%), and four had had a spontaneous aortoduodenal fistula (1%). In all these patients resections were undertaken, but in another 56 patients (11%) the aneurysm was not resected. A review of these cases showed that 91% had symptoms at their first presentation; abdominal pain and backache being most common. The diagnosis could be established in 91% by the presence of pulsatile abdominal mass on clinical examination. The operative mortality for elective resection was 8%, for acute 19%, for ruptured cases 42%, and for spontaneous aortoduodenal fistula 50%. After successful resection the overall five-year survival was 65% by the life table method, and there was no significant difference between elective, acute, and ruptured cases. This five-year survival after resection compares favourably with the expected 76% survival of a similar normal population, and was considerably better than that for conservatively treated patients. As most cases have symptoms, and diagnosis may be established easily by routine physical examination in 91%, the prognosis for this condition could be considerably improved by increased awareness of its existence and early referral for treatment as an elective surgical procedure.